Evaluation of cell lines and immunofluorescence and plaque assay procedures for quantifying reoviruses in sewage

Shin

Microbiology and Immunology

2006

Contaminated water plays a key role in the transmission of enteric viruses because water is the main route by which these viruses spread in a population. Enteric viruses are transmitted in high numbers via the fecaloral route and are closely related with waterborne viral diseases. Enteric viruses have been found in rivers, groundwater, and even in treated drinking water (1, 6) and are recognized as major public health hazards. They are known to cause various human illnesses such as meningitis, encephalitis, hepatitis, respiratory illnesses, gastroenteritis, and skin rashes, and include the polioviruses, coxsackieviruses, rotaviruses, hepatitis A virus (HAV), noroviruses, adenoviruses, and reoviruses (3,8,14).The common mammalian reovirus isolates are type 1 (Lang), type 2 (Jones), and type 3 (Dearing and Abney) and have been isolated from children with diarrhea or upper respiratory infections. Reoviruses are known to have a high endemic infection rate among humans and other animals. They typically cause an asymptomatic or mild respiratory infection and gastroenteritis. Due to the resistance of reoviruses to chlorination and their high stability in water, these viruses can induce severe bacterial respiratory disease after initial reovirus infection especially in the immunocompromised, young, and old individuals (16). Many studies have examined environmental water samples for enteric viruses (5,12,13,15,17). While most virus-monitoring methods mainly target enteroviruses in water samples, we targeted reoviruses as well as other enteric viruses, including adenoviruses and HAV (20,21). Reoviruses are known to be present in water in greater numbers than enteroviruses, and have been frequently detected in surface waters either alone or associated with enteroviruses (22).The total culturable virus assay (TCVA) has been widely used as a standard method for the detection and quantification of infectious enteric viruses from environmental waters. A PCR-based assay is a powerful technique for detecting target RNA or DNA and could reduce the cost and time, and increase the detection sensitivity of a monitoring program (1, 9). However, a Detection of Enteroviruses and Mammalian Reoviruses in Korean Environmental

Section: Methodsmentioning

confidence: 99%

Detection of Enteroviruses and Mammalian Reoviruses in Korean Environmental Waters

Shin

Microbiology and Immunology

2006

International Journal of Hygiene and Environmental Health

“…Smith and Gerba (1982) have shown that as few as 10 infectious units per liter can be reliably detected by IFA within less than 24 h incubation. Ridinger et al (1982) found that IFA is more sensitive and more rapid than the plaque assay for the detection of reoviruses. Reoviruses in excess of 10 4 per liter of raw sewage were detected by IFA.…”

Section: Fluorescence Microscopy and Virus Infectivitymentioning

confidence: 98%

“…This method uses virus-specific antibody labelled with a fluorescent dye to visualize cell-associated viral antigens (Jeffery and Aarons, 2009). The use of the IFAs has been described previously for detecting infectious rotaviruses in sewage (Ridinger et al, 1982;Smith and Gerba, 1982). Smith and Gerba (1982) have shown that as few as 10 infectious units per liter can be reliably detected by IFA within less than 24 h incubation.…”

Section: Fluorescence Microscopy and Virus Infectivitymentioning

confidence: 99%

Methods to detect infectious human enteric viruses in environmental water samples

Hamza

Jurzik

Überla

et al. 2011

131

Currently, a wide range of analytical methods is available for virus detection in environmental water samples. Molecular methods such as polymerase chain reaction (PCR) and quantitative real time PCR (qPCR) have the highest sensitivity and specificity to investigate virus contamination in water, so they are the most commonly used in environmental virology. Despite great sensitivity of PCR, the main limitation is the lack of the correlation between the detected viral genome and viral infectivity, which limits conclusions regarding the significance for public health. To provide information about the infectivity of the detected viruses, cultivation on animal cell culture is the gold standard. However, cell culture infectivity assays are laborious, time consuming and costly. Also, not all viruses are able to produce cytopathic effect and viruses such as human noroviruses have no available cell line for propagation. In this brief review, we present a summary and critical evaluation of different approaches that have been recently proposed to overcome limitations of the traditional cell culture assay and PCR assay such as integrated cell culture-PCR, detection of genome integrity, detection of capsid integrity, and measurement of oxidative damages on viral capsid protein. Techniques for rapid detection of infectious viruses such as fluorescence microscopy and automated flow cytometry have also been suggested to assess virus infectivity in water samples.

Cytokine & Growth Factor Reviews

“…Reovirus (respiratory enteric orphan virus) is a member of the Reoviridae family. Reovirus is ubiquitous and has been isolated from untreated sewage, stagnant water and rivers throughout the world [21,22]. As the name suggests, reovirus infects the respiratory and gastrointestinal tract but is not associated with a named human disease, although it can infrequently cause flu-like upper respiratory tract symptoms or a mild diarrhoeal illness.…”

Section: Oncolytic Reovirus As An Archetype For Clinical Developmentmentioning

confidence: 99%

Clinical trials with oncolytic reovirus: Moving beyond phase I into combinations with standard therapeutics

Harrington

Vile

Melcher³

et al. 2010

It is time for those working on oncolytic viruses to take stock of the status of the field. We now have at our disposal an array of potential therapeutic agents, and are beginning to conduct early-phase clinical trials in patients with relapsed/metastatic cancers. By drawing on lessons learned during the development of other biological therapies, such as monoclonal antibodies and targeted small molecule inhibitors, we are now in a position to chart the course of the next wave of trials that will go beyond the phase I studies of safety and feasibility. In this article we review our approach to the development of oncolytic viruses as cancer therapeutics. In doing so, we emphasise the fact that this process is modular and involves multiple iterative steps between the laboratory and the clinic. Ultimately, at least in the medium term, the future of oncolytic virotherapy lies in combination regimens with standard anti-cancer agents such as radiation and chemotherapy.